The study of the pathway of clavine alkaloid biosynthesis and the enzymes involved in the pathway will continue. Fluorometric assay procedure will be developed for gamma, gamma-4-dimethylallyltryptophan-oxygen oxidoreductase (forming clavicipitic acid, hydrogen peroxide) (DMAT oxidase). The enzyme will be further purified for kinetic and other mechanistic studies. The chemical synthesis of allylic alcohol derivatives of DMAT is in progress. The first step in the synthesis is the oxidation of dimethylallylindole with osmium tetroxide. N alpha-(C3H3) methyl (sidechain 3-C14) DMAT will be synthesized and tested as a precursor of the clavine alkaloids. The conversion of elymoclavine to agroclavine observed in cultures of Claviceps purpurea PRL 1980 is being investigated. To test the possible role of dihydrolysergol in the conversion, radioactive dihydrolysergol will be synthesized by reduction of elymoclavine and tested for incorporation into agrovlavine. The enzymatic exchange of a hydrogen in DMAT will be studied. The location of deuterium in exchanged DMAT will be located by mass spectrometry. The properties of the exchange reaction will be studied. BIBLIOGRAPHIC REFERENCES: Bajwa, R.S., and Anderson, J.A., Conversion of Agroclavine to Setoclavine and Isosetoclavine in Cell-free Extracts from Claviceps sp. DS 58 and in a Thioglycolate/Iron (II) System. J. Pharm. Sci., 64: 343-344 (1975). Bajwa, R.S., Kohler, R.D., Cheng, M., Saini, M.S., and Anderson, J.A., Formation and Clavicipitic Acid from 4-Dimethylallyltryptophan in Cell-free Systems of Claviceps sp. SD 58, Phytochem., 14: 735-737 (1975).